康莱特注射液对HL-60/ADR细胞周期调控蛋白P16、P63表达的影响

The Effect of Kanglaite Injection on Expression of Cyclin Modulin P16, P63 in HL-60/ADR Cells

关键词:  康莱特注射液  HL-60/ADR细胞  细胞周期  P16  P63

Key words:  Kanglaite injection  HL-60/ADR cells  cell cycle  P16  P63

摘要:

目的 探索康莱特注射液对HL-60/ADR细胞周期调控蛋白P16、P63表达的影响。方法 使用康莱特注射液作用于HL-60/ADR细胞，用细胞计数法检测康莱特注射液对HL-60细胞生长曲线的影响，MTT法检测阿霉素对康莱特注射液作用后细胞的抑制效应。康莱特注射液作用HL-60/ADR细胞一定时间后，再用一定浓度的阿霉素作用该细胞一定时间，流式细胞仪检测该细胞内的阿霉素浓度并分析细胞周期调控蛋白P16、P63的表达。结果 各组细胞计数第6天达峰值，康莱特注射液各组细胞计数，较未加药组细胞计数低(P<0.05)。1 mg·mL^-1康莱特注射液作用HL-60/ADR细胞96 h，阿霉素对该细胞的抑制效应较对康莱特注射液未作用的HL-60/ADR细胞抑制效应强(P<0.05)；不同浓度的康莱特注射液作用HL-60/ADR细胞96 h后，再用1.5 μg·mL^-1的阿霉素作用6 h，该细胞内的阿霉素荧光强度平均值(127.2±4.8)较对照组(77.8±7.2)高(P<0.05)，且具有剂量依赖关系；1 mg·mL^-1康莱特注射液作用HL-60/ADR细胞48 h，HL-60/ADR细胞的细胞周期调控P16的平均值(898.46±30.50)较对照组(698.45±91.29)高(P<0.05)；P63的平均值(837.53±42.75)较对照组(591.33±92.99)高(P<0.05)。结论 康莱特注射液对HL-60细胞生长有抑制作用，可能有促进细胞对阿霉素的敏感性的作用。康莱特注射液还有上调HL-60/ADR细胞周期调控蛋白P16、P63的表达，此机制可能参与康莱特注射液抗肿瘤作用。

Abstract:

OBJECTIVE To investigate the effect of Chinese medicine Kanglaite injection on expression of cyclin modulin P16, P63 in HL-60/ADR cells. METHODS HL-60/ADR cells during logarithmic growth phase were treated with Kanglaite injection, growth curve was based on cell counting; the inhibitory effect of adriamycin(ADR) on Kanglaite injection-treated HL-60/ADR cells was tested by MTT assay; the concentration of intracellular adriamycin and the expression of P16, P63 were detected by flow cytometry. RESULTS In each group, live cell count reached a peak on the 6th day, compared with control group, the Kanglaite injection-treated cell count was lower with statistical significance(P<0.05). Treated HL-60/ADR cells with 1 mg·mL^-1 Kamglaite for 96 h, the inhibitory effect of adriamycin on the cells were greater than that of control groups(P<0.05). After treatment with different concentration of Kanglaite injection, the intrancellular ADR fiourescence of 1.5 μg·mL^-1 ADR-treated for 6 h were statistical higher than that of control group(P<0.05). Treated HL-60/ADR cells with 1 mg·mL^-1 Kamglaite for 48 h, compared with the control group, the expression of P16, P63 were all significantly increased(P<0.05). CONCLUSION HL-60/ADR cells exposed to Kanglaite injection resulted in dose-depentent inhibitory, Kanglaite injection could increase the sensitivity of HL-60/ADR cells to ADR. Kanglaite injection could increase the expression level of P16, P63 in HL-60/ADR cells, that maybe involved in the anti-tumor mechanics of Kanglaite injection.